FAQ: What primers should I use to sequence the ends of my insert after I clone it into a pMAL vector?
The following sequencing primers can be used (not available from New England Biolabs): malE primer on the 5′ side of the insert and the pMAL reverse primer for sequencing from the 3′ side.
Forward Primer (24-mer, upstream of MCS)
5′d(GGTCGTCAGACTGTCGATGAAGCC)3′
Reverse Primer (24-mer, downstream of MCS)
5′d(TGTCCTACTCAGGAGAGCGTTCAC)3′
The sequences of the pMAL vectors are also available at www.neb.com. PCR of inserts cloned into the MCS can be performed using the same primer pair.